title: In planta protein interactions of three alphacryptoviruses and three betacryptoviruses from White Clover, Red Clover and Dill by bimolecular fluorescence complementation analysis.
abstract: Plant-infecting viruses of the genera Alpha- and Betacryptovirus within the family Partitiviridae cause no visible effects on their hosts and are only transmitted by cell division and through gametes. The bipartite dsRNA genome is encoding a RNA-dependent RNA polymerase (RdRp) and a coat protein (CP). Aside from sequence and structural analysis, the investigation of protein interactions is another step towards virus characterization. Therefore, ORFs of two type members White Clover Cryptic Virus 1 and 2 (WCCV-1 and WCCV-2), as well as the related viruses from Red Clover and Dill were introduced into a bimolecular fluorescence complementation assay. We showed CP-CP dimerization for all tested viruses with localization for alphacryptoviruses at the nuclear membrane and for betacryptoviruses close to cell walls within the cytoplasm. For CPs of WCCV-1 and WCCV-2, deletion mutants were created to determine internal interaction sites. Moreover, RdRp self-interaction was found for all viruses, whereas CP-RdRp interactions were only detectable for the alphacryptoviruses. An intra-genus test of CPs was successful in various virus combinations, whereas an inter-genus interaction of WCCV-1CP and WCCV-2CP was absent. This is the first report of in vivo protein interactions of members in the family Partitiviridae, indicating distinct features of the alpha- and betacryptoviruses.
authors.affiliation: Department of Phytomedicine, Institute of Horticultural Production Systems, Leibniz Universität Hannover, Herrenhäuser Str. 2, Hannover D-30419, Germany. maiss@ipp.uni-hannover.de.
authors.forename: Till
authors.initials: T
authors.lastname: Lesker
authors.forename: Edgar
authors.initials: E
authors.lastname: Maiss
mesh_terms: D028524:Anethum graveolens / Q000821:virology*; D005453:Fluorescence; D008969:Molecular Sequence Data; D010942:Plant Viruses / Q000235:genetics / Q000502:physiology*; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D012328:RNA Viruses / Q000235:genetics / Q000502:physiology*; D012367:RNA, Viral / Q000235:genetics; D017422:Sequence Analysis, DNA; D029921:Trifolium / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins